Chapter

The Textbook of Angiogenesis and Lymphangiogenesis: Methods and Applications

pp 217-240

Date:

Visualization and Quantification of De Novo Angiogenesis in Ex Ovo Chicken Embryos

  • A. ZijlstraAffiliated withDepartment of Pathology, Vanderbilt University
  • , John D. LewisAffiliated withDepartment of Oncology, University of Alberta Email author 

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Abstract

The formation of new blood vessels through de novo angiogenesis is a fundamental process in developmental biology and pathogenesis, during which changes in the vasculature can be highly dynamic and involve proliferation, migration, sprouting, and remodeling. In this chapter, methodologies are described which allow for the precise temporal and spatial quantification of angiogenesis in an ex ovo chicken embryo model. A protocol is detailed that allows for the enumeration of blood vessels in newly vascularized collagen onplants placed on the chorioallantoic membrane. Onplants can be infused with growth factors, cytokines, pro- or anti-angiogenic factors or even living cells to determine the precise impact on de novo angiogenesis. A complementary intravital imaging approach is also described that allows for the visualization and evaluation of angiogenesis in ex ovo chicken embryos as it occurs in real time. These highly scalable assays evaluate new vessel growth in an intact tissue where angiogenesis naturally occurs, and importantly, allow one to evaluate factors that inhibit vascularization without impacting pre-existing vessels.